Part:BBa_K3490010
Verifying the function of tetR system.
Our first kill switch design uses TetR fused with VP16, tet-off system[1]to control lysozyme as our biosafety to prevent our engineered bacteria surviving when the contact lens accidently breaks (BBa_K3490011). (Fig.1)
Fig. 1 Our design about the killswitch using tet system
After trying many times to transform our IDT synthesis into E. coli, we failed as precedent because VP16 hasn’t been applied to prokaryotic. To prove the concept that bacteria will die when escaping the contact lens, we found the plasmid from iGEM kit BBa_K611059, which has pTet promoter with GFP, and pBAD promoter with TetR. We planned to use AnhydroTetracycline to do the GFP express experiment[2], but it is too expensive to do it, so we found another way to prove our ideas. When arabinose and heat inactivate chlortetracycline (CTC) [2] or Tetracycline (Tc) is added into the liquid culture, it should express a different amount of GFP, as to proof heat inactivate chlortetracycline or Tetracycline is crucial to the kill switch system.
Still in progress......
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 755
Illegal NheI site found at 1068
Illegal NheI site found at 1091 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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